| Image | Western BlotPositive WB detected in: 3 different overexpression lysates with HA taggedAll lanes: HA-Tag antibody at 1:1000SecondaryGoat polyclonal to Mouse IgG at 1/10000 dilutionPredicted band size: 35, 35, 48 kDaObserved band size: 35, 35, 48 kDa Western BlotPositive WB detected in: HA-tagged fusion protein at 50ng, 25ng, 12.5ng, 6.25ng, 3.125ng, 1.5625ngAll lanes: HA-Tag antibody at 1:1000SecondaryGoat polyclonal to Mouse IgG at 1/10000 dilutionPredicted band size: 35 kDaObserved band size: 35 kDa Western BlotPositive WB detected in: HA-tagged fusion proteinAll lanes: HA-Tag antibody at 1:5000, 1:10000, 1:20000, 1:40000, 1:80000, 1:160000SecondaryGoat polyclonal to Mouse IgG at 1/10000 dilutionPredicted band size: 35 kDaObserved band size: 35 kDa Western BlotPositive WB detected in: 6 different recombinant proteins with HA tagged, Hela whole cell lysate, 3T3 whole cell lysateAll lanes: HA-Tag antibody at 1:5000SecondaryGoat polyclonal to Mouse IgG at 1/10000 dilution Immunofluorescence staining of 293F cells with CSB-MA000141M0m at 1:100, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L). Immunofluorescence staining of 293F transfected cells with CSB-MA000141M0m at 1:100, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L). Immunofluorescence staining of 293F cells and 293F transfected cells with Company A, Company B, Company C, Company D, Company E, CSB-MA000141M0m at 1:100, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L). Immunoprecipitating HA-Tag in 293F transfected whole cell lysateLane 1: Mouse control IgG (1µg) instead of CSB-MA000141M0m in 293F transfected whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)Lane 2: CSB-MA000141M0m (5µg) + 293F transfected whole cell lysate (500µg)Lane 3: 293F transfected whole cell lysate (20µg) Immunoprecipitating HA-Tag in 293F transfected whole cell lysateLane 1: Mouse control IgG (1µg) instead of CSB-MA000141M0m in 293F transfected whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)Lane 2: Company A (5µg), Company B (5µg),Company C (5µg),Company D (5µg),Company E (5µg),CSB-MA009476A0m (5µg) + 293F transfected whole cell lysate (500µg)Lane 3: 293F transfected whole cell lysate (20µg) Overlay histogram showing 293F transfected cells stained with CSB-MA000141M0m (red line) at 1:100. The cells were incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.
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